Hair lysate and application thereof

Abstract

The invention relates to a hair lysate and application thereof. The hair lysate is prepared from the following components in percentage by volume: 0.05 to 1.0 percent of ethyl phenyl polyethylene glycol, 2 to 18 mg / mL of dithiothreitol and 2 to 20 U / mL of protease. The NP-40 is a surfactant, the dithiothreitol is a reducing agent, and the NP-40 and the dithiothreitol jointly act with the protease, so that drug components possibly existing in the hair can be quickly released, ultrasonic or freezing grinding is not needed, and the operation is simple and convenient. The hair lysate can meet the requirement of qualitative detection only after being treated for 10 minutes, the best effect can be obtained if the treatment time is as long as 30 minutes, the release efficiency is superior to that of a traditional method, and whether a hair sample contains trace drugs or not can be accurately detected by matching with subsequent liquid chromatography-tandem mass spectrometry analysis.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of sample pretreatment technology, in particular to a hair lysis solution and application thereof. BACKGROUND

[0002] Drug abuse refers to the repeated use of some drugs with dependency or potential dependency for non-medical purposes, which is self-administered by the user, leading to mental and physical dependence, causing mental confusion and abnormal behavior. Drug abuse is also known as "drug use" or "drug abuse". Drug use is deeply related to family breakdown, violent crime, economic poverty and other problems, and poses a serious challenge to social stability and public health.

[0003] Detecting psychotropic drug residues in biological samples is an important basis for public security and judicial departments to determine whether the sample provider has drug use behavior. In the field of drug detection, hair drug detection technology has become a research hotspot in forensic science and clinical analysis due to its unique advantages. After drugs enter the human body, they circulate through the blood into the hair follicle, and the drug molecules and their metabolites are fixed and stably retained in the keratin protein of the hair, which makes it possible to trace drug use behavior over a long period of time. Compared with urine, blood, saliva and other biological sample detection technologies, hair drug detection has a longer tracing period, usually 3 to 6 months of drug abuse behavior, and if the sample provider has long hair, the tracing range can even extend to several years ago. In addition, hair drug detection has a wide sampling range, not limited to head hair, and body hair such as pubic hair, armpit hair, and nose hair can also be used for detection, and some hair can detect more types of drugs with more accurate content. Hair detection is unique, similar to DNA detection, and does not require collateral evidence. If the detection result is positive and the person being tested has a drug record, it can be determined that the person has used drugs. It has a significant effect on identifying hidden drug users, monitoring the effectiveness of drug rehabilitation, identifying drug addiction, and deterring various high-risk and sensitive drug-related groups.

[0004] Currently, sample pretreatment is required before hair drug detection, and the main treatment methods include organic solvent ultrasonic method, acid hydrolysis, alkaline hydrolysis, and enzymatic hydrolysis, and techniques such as immunocolloidal gold technology, gas chromatography-mass spectrometry analysis, and high-performance liquid chromatography-mass spectrometry are used for detection and analysis. Researchers have established a series of accurate, rapid, and highly sensitive qualitative and quantitative analysis methods to detect new drugs according to different application scenarios.

[0005] In hair pretreatment technology, the organic solvent ultrasonic method generally uses solvents such as methanol, ethanol, and acetonitrile to extract drug molecules from hair with ultrasonic oscillation, but this method has the problems of complex operation, long extraction time, and incomplete release. Acid hydrolysis has a mild condition, a wide range of applications, and a relatively simple operation, but it is time-consuming and not suitable for some drug molecules (such as cocaine, O 6The release rate of monoacetylmorphine is low. The alkaline hydrolysis method is a severe condition that can completely digest and dissolve the hair, so that the amphetamines in the hair are fully released, but there are many impurities, and the compounds with unstable structures are prone to degradation under strong alkaline conditions. The enzymatic reaction is concerned in the pre-treatment of hair drug detection due to its mild conditions and strong specificity. Some products on the market for detecting drugs in hair all claim to have the characteristics of fast time efficiency, good safety, and low cost, but the actual range of detectable drug types is small, the detection specificity is poor, and false negatives, false positives, weak negatives, and weak positives are easy to occur. The currently disclosed cracking solution formula contains high-concentration sodium salt components (such as NaCl, NaOH, and sodium sulfite), which can significantly increase the ionic strength of the solution system from the principle of analytical chemistry. In the liquid chromatography-mass spectrometry (LC-MS / MS) analysis based on electrospray ionization (ESI), the high ionic strength will cause ion suppression effect, resulting in a significant decrease in the ionization efficiency of the target analyte. The specific performance is that the ionization process of the analyte on the droplet surface is blocked, the number of ions entering the gas phase is reduced, and finally the liquid chromatography-mass spectrometry response signal is significantly weakened, which seriously affects the detection sensitivity and accuracy. In view of the above technical bottlenecks, the cracking solution containing high-concentration sodium salt is not suitable for direct analysis of LC-MS / MS, so it is often necessary to replace the traditional liquid chromatography-mass spectrometry detection method for re-measurement and verification.

[0006] In the current field of hair drug detection, the traditional liquid chromatography-mass spectrometry detection method uses frozen grinding and ultrasonic extraction. SF / ZJD0107025-2018 "Liquid chromatography-tandem mass spectrometry detection method for 15 drugs and metabolites in hair" is a commonly used detection standard. According to the standard, the process of hair drug detection covers multiple detailed steps. First, the hair sample needs to be frozen and ground, which usually takes about 30 minutes, followed by ice bath ultrasonic treatment, also lasting for 30 minutes, then centrifugation, which takes about 10 minutes, and finally blow-drying and re-dissolving, which generally takes 30 minutes. From the overall process, the traditional hair drug detection method according to this standard takes about 2 hours for the entire pre-treatment process. Such a long pre-treatment time not only reduces the detection efficiency, but also causes sample backlog in actual application scenarios, such as large-scale drug screening, affecting the timely feedback of the detection results. Moreover, the process involves multiple complex steps, and the professional skills of the experimenters are required, and any deviation in any step may affect the final detection results, making the operation process complex. SUMMARY

[0007] The present application aims to disclose a hair cracking solution and its application, to solve one or more technical problems in the prior art, and to provide at least one beneficial option or create conditions.

[0008] The first aspect of the present application is to provide a hair lysis solution.

[0009] The second aspect of the present application is to provide the use of the hair lysis solution according to the first aspect of the present application.

[0010] The hair lysis solution according to the first aspect of the present application comprises 0.05-1.0% (v / v) NP-40, 2-18 mg / mL DTT and 2-20 U / mL protease. The NP-40 is a surfactant, and the DTT is a reducing agent. The possible drug components in the hair can be quickly released by the joint action of the protease, without the need for ultrasonic or cryogenic grinding, and the operation is simple. The hair lysis solution can meet the requirements of qualitative detection after 10 minutes of treatment. If the treatment lasts for 30 minutes, the best effect can be obtained, and the release efficiency is better than that of the traditional method. Moreover, the hair lysis solution can be combined with subsequent liquid chromatography-tandem mass spectrometry (LC-MS / MS) analysis to accurately detect whether the hair sample contains trace amounts of drugs.

[0011] In a further application embodiment, the hair lysis solution comprises 0.1-0.5% (v / v) NP-40, 2-10 mg / mL DTT and 4-10 U / mL protease.

[0012] In a further application embodiment, the hair lysis solution further comprises a buffer selected from a phosphate buffer, a Tris-HCl buffer or a citric acid buffer. Preferably, the buffer is a Tris-HCl buffer. This is because the Tris-HCl buffer has a pH of 7.4, and the DTT is relatively stable in the pH range of 7-8, which can effectively prolong the shelf life of the hair lysis solution.

[0013] In a further application embodiment, the hair lysis solution further comprises 0.01-0.3% (v / v) β-mercaptoethanol (β-ME). The β-ME is both a reducing agent and an antioxidant, which can prevent the oxidation of the easily oxidizable groups in the DTT and the protease, prevent inactivation and improve stability, prolong the shelf life of the hair lysis solution and ensure the lysis effect.

[0014] In a further application embodiment, the hair lysis solution further comprises 10-40 mg / mL of sodium dodecyl sulfate (SDS). SDS loosens the hair structure by breaking the non-covalent bonds of the hair keratin, helping the protease to fully contact and improving the enzymatic efficiency and detection sensitivity. However, as a cationic surfactant, SDS has strong ionization ability, so when the SDS component is added to the hair lysis solution, an extraction and purification step needs to be added after the constant temperature reaction to eliminate the interference of SDS on LC-MS / MS.

[0015] In a further application embodiment, the protease comprises protease K and / or keratinase. The main component of hair is keratin. The keratin structure is tight, and ordinary proteases are difficult to decompose, while protease K and keratinase can specifically recognize and cut specific peptide bonds in keratin, decomposing it into smaller peptide segments and amino acids, thereby achieving rapid enzymatic hydrolysis of hair.

[0016] In a further application embodiment, the hair lysis solution formula includes scheme one and scheme two. Scheme one is suitable for direct filtration of the sample after constant temperature reaction, and LC-MS / MS detection can be performed; scheme two contains SDS, and the interference of SDS needs to be removed by liquid-liquid extraction method after constant temperature reaction, and then LC-MS / MS detection is performed. Scheme one is suitable for rapid extraction and detection of etomidate, propylpiperidolate, isopropylpiperidolate, morphine, O 6 - rapid extraction and detection of single acetyl morphine, codeine, methamphetamine, amphetamine, ketamine, norketamine, MDMA, MDA, MDEA, etc. Scheme two can further detect cannabis and synthetic cannabinoid drugs based on scheme one.

[0017] The second aspect of the present application provides the use of the hair lysis solution of the first aspect of the present application. Specifically, it is used for pretreating human hair, so that the treated sample can be used for LC-MS / MS to detect trace drugs.

[0018] In a further application embodiment, the detection comprises the following steps: S1. According to the "Hair Sample Detection Standard for Drug Involved Personnel", collect 20 mg of hair about 3 cm from the root end, cut into small pieces of 0.5-2 mm, add the hair lysis solution, and vortex mix; S2. Perform constant temperature reaction; S3. Filter and perform liquid chromatography-tandem mass spectrometry analysis.

[0019] In a further application embodiment, 5-20 mg of cut hair is weighed in S1, and 50±5 μL of the hair lysis solution is added per milligram of hair sample.

[0020] In a further application embodiment, the constant temperature reaction has a temperature of 25-65°C.

[0021] In a further application embodiment, the constant temperature reaction has a reaction time of 10-30 minutes.

[0022] In a further application embodiment, for the second scheme of the hair lysis solution, step S2 further comprises adding chloroform and isopropyl alcohol to the constant temperature reaction product for extraction, centrifuging to obtain the organic phase, blowing dry, and then redissolving in methanol. The components in the first scheme of the hair lysis solution do not have obvious ionization inhibition effect on most types of drugs in LC-MS / MS detection, but the cannabinoids and synthetic cannabinoids will compete for charges with the hair lysis solution during ionization, resulting in reduced ionization efficiency of the target substances and ion inhibition effect. Therefore, SDS is added as the second scheme on the basis of the first scheme, and a mixed solution of chloroform and isopropyl alcohol is used to extract and purify the small molecule drugs in the constant temperature reaction product, so as to reduce the inhibition effect of the interfering substances on cannabinoids and synthetic cannabinoids, and improve the mass spectrometry detection sensitivity of this type of drugs.

[0023] In a further application embodiment, the chromatographic conditions of the liquid chromatography-tandem mass spectrometry analysis are as follows: The chromatographic column is a Restek Allure PFP Propyl chromatographic column with a column length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 5 μm; The mobile phase is 5 mmol / L ammonium acetate and 0.1% formic acid buffer, and acetonitrile gradient elution; The mass spectrometry conditions are electrospray ionization positive ion mode and multiple reaction monitoring. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 The detection spectrum of the morphine and codeine positive sample in Example 1; Figure 2 The detection spectrum of the ketamine positive sample in Example 2; Figure 3 The detection spectrum of the methamphetamine positive sample in Example 3; Figure 4 The detection spectrum of the etomidate positive sample in Example 4; Figure 5 The detection spectrum of the nimetazepam positive sample in Example 5; Figure 6 The detection spectrum of the clonazepam positive sample in Example 6; Figure 7 The detection spectrum of the negative sample in Example 7; Figure 8Figure 1 shows a control chromatogram for the detection of a cannabinol positive sample using protocol one and protocol two of the hair lysis solution used in Example 8. Figure 9 Figure 2 shows a fast screening test report for a commercially available product positive sample tested in Example 10. DETAILED DESCRIPTION

[0025] In the description of the application, if there is a description of first, second, etc., it is only for the purpose of distinguishing technical features, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features or the sequence of indicated technical features.

[0026] The words "preferably", "more preferably" and the like in the present invention refer to embodiments of the present invention that can provide certain benefits under certain circumstances. However, other embodiments can also be preferred under the same or other circumstances. In addition, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the present invention.

[0027] When a numerical range is disclosed herein, the range is to be construed as continuous, and includes the minimum and maximum values of the range, as well as every value between the minimum and maximum values. Further, when ranges are provided for integers, every integer between the minimum and maximum values is included. In addition, when multiple ranges are provided to describe a characteristic or property, the ranges can be combined. In other words, unless otherwise indicated, all ranges disclosed herein are to be understood as including any and all sub-ranges subsumed therein.

[0028] The technical solutions in the embodiments of the present application will be described clearly and completely below in conjunction with the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, not all. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of the present application.

[0029] The reagents, methods and equipment used in the present application are conventional reagents, methods and equipment in the technical field, unless otherwise specified.

[0030] The hair lysis solution protocol one is configured as follows: 0.1% (volume percentage) NP-40, 0.05% (volume percentage) β-ME, 6 mg DTT, and 5 U protease K are contained in 1 mL of 0.5 mol / L Tris-HCl buffer (pH=7.4).

[0031] The hair lysis solution scheme two is configured in the following manner: in 1 mL of 0.5 mol / L Tris-HCl buffer (pH=7.4), the following components are contained: NP-40 content 0.1% (volume percentage), β-ME content 0.05% (volume percentage), DTT content 6 mg, proteinase K content 5 U, and SDS content 20 mg.

[0032] Example 1, detection of morphine positive sample The hair sample of the suspected drug user confirmed by the law inspection as positive for morphine is obtained, and the hair lysis solution of scheme one is used for rapid detection, and the specific operation steps are as follows: S1. The hair sample is cut to 0.5-2 mm, 50±5 μL of the hair lysis solution is added to 1 mg of hair, and vortexed for uniform mixing; S2. 37℃ constant temperature reaction for 15 minutes; S3. Filter the lysis solution and perform LC-MS / MS analysis.

[0033] The hair lysis solution of scheme two is used for detection, and the specific operation steps are as follows: S1. The hair sample is cut to 0.5-2 mm, 50±5 μL of the hair lysis solution is added to 1 mg of hair, and vortexed for uniform mixing; S2. 37℃ constant temperature reaction for 15 minutes, a mixed solution of chloroform and isopropanol is added to the reaction product for extraction, and after centrifugation, the organic phase is blown dry in air flow, and redissolved with methanol; S3. Filter the sample solution and perform LC-MS / MS analysis.

[0034] The LC-MS / MS detection results are shown in Figure 1 It is shown that using the hair lysis solution of scheme one and scheme two and LC-MS / MS detection can accurately identify morphine and codeine in the hair of the suspected drug user as positive, without impurity interference, which is consistent with the law inspection results.

[0035] Example 2, detection of ketamine positive sample The hair sample of the suspected drug user confirmed by the law inspection as positive for ketamine is obtained, and the hair lysis solution of scheme one is used for rapid detection, and the specific operation steps are as follows: S1. The hair sample is cut to 0.5-2 mm, 50±5 μL of the hair lysis solution is added to 1 mg of hair, and vortexed for uniform mixing; S2. 37℃ constant temperature reaction for 15 minutes; S3. Filter the lysis solution and perform LC-MS / MS analysis.

[0036] The LC-MS / MS detection results are shown in Figure 2As shown in Table 1, it is shown that using the hair lysate and LC-MS / MS detection, the ketamine in the hair of the drug-involved person can be accurately determined to be positive without impurity interference, which is consistent with the result of the law inspection.

[0037] Example 3, detection of positive samples of new psychoactive drugs of etomidate Hair samples of drug-involved persons confirmed by law inspection to be positive for new psychoactive drugs of etomidate were obtained, and the hair lysate of scheme one was used for rapid detection, and the specific operation steps were as follows: S1. Cut the hair sample to 0.5-2 mm, add 50±5 μL of the hair lysate to 1 mg of hair, and vortex mix; S2. Incubate at 37°C for 15 minutes; S3. Filter the lysate and perform LC-MS / MS analysis.

[0038] The LC-MS / MS detection results are shown in Table 2, which show that using the hair lysate and LC-MS / MS detection, the etomidate in the hair of the drug-involved person can be accurately determined to be positive without impurity interference, which is consistent with the result of the law inspection. Figure 3

[0039] Example 4, detection of methamphetamine positive samples Hair samples of drug-involved persons confirmed by law inspection to be positive for methamphetamine were obtained, and the hair lysate of scheme one was used for rapid detection, and the specific operation steps were as follows: S1. Cut the hair sample to 0.5-2 mm, add 50±5 μL of the hair lysate to 1 mg of hair, and vortex mix; S2. Incubate at 37°C for 15 minutes; S3. Filter the lysate and perform LC-MS / MS analysis.

[0040] The hair lysate of scheme two was used for detection, and the specific operation steps were as follows: S1. Cut the hair sample to 0.5-2 mm, add 50±5 μL of the hair lysate to 1 mg of hair, and vortex mix; S2. Incubate at 37°C for 15 minutes, add a mixed solution of chloroform and isopropyl alcohol to the reaction product, centrifuge, and take the organic phase to blow dry in air flow, and redissolve with methanol; S3. Filter the sample solution and perform LC-MS / MS analysis.

[0041] The LC-MS / MS detection results are shown in Table 3, which show that using the hair lysate of scheme one and scheme two and LC-MS / MS detection, the methamphetamine in the hair of the drug-involved person can be accurately determined to be positive without impurity interference, which is consistent with the result of the law inspection. Figure 4 LC-MS / MS detection results are shown in Table 3, which show that using the hair lysate of scheme one and scheme two and LC-MS / MS detection, the methamphetamine in the hair of the drug-involved person can be accurately determined to be positive without impurity interference, which is consistent with the result of the law inspection.​

[0042] Example 5, detection of positive sample of nimetazepam Hair samples of suspected drug addicts confirmed positive for nimetazepam by forensic examination were obtained and detected using the hair lysis solution of Scheme II, with the following specific steps: S1. Cut the hair sample into 0.5-2 mm, add 50±5 μL of the hair lysis solution to 1 mg of hair, and vortex mix; S2. React at 37°C for 15 minutes, add a mixed solution of chloroform and isopropanol to the reaction product for extraction, and after centrifugation, take the organic phase and dry it in air flow, and redissolve it in methanol; S3. Filter the sample solution and perform LC-MS / MS analysis.

[0043] The LC-MS / MS detection results are shown in Figure 5 , which show that using the hair lysis solution and LC-MS / MS detection can accurately determine the positive nimetazepam in the hair of drug addicts, without impurity interference, consistent with the forensic results.

[0044] Example 6, detection of positive sample of clonazepam Hair samples of suspected drug addicts confirmed positive for clonazepam by forensic examination were obtained and detected using the hair lysis solution of Scheme II, with the following specific steps: S1. Cut the hair sample into 0.5-2 mm, add 50±5 μL of the hair lysis solution to 1 mg of hair, and vortex mix; S2. React at 37°C for 15 minutes, add a mixed solution of chloroform and isopropanol to the reaction product for extraction, and after centrifugation, take the organic phase and dry it in air flow, and redissolve it in methanol; S3. Filter the sample solution and perform LC-MS / MS analysis.

[0045] The LC-MS / MS detection results are shown in Figure 6 , which show that using the hair lysis solution and LC-MS / MS detection can accurately determine the positive clonazepam in the hair of drug addicts, without impurity interference, consistent with the forensic results.

[0046] Example 7, detection of negative sample Hair samples of people who did not use prohibited drugs were obtained and detected using the hair lysis solution of Scheme I and Scheme II, respectively.

[0047] The detection results are shown in Figure 7 , which show that the corresponding retention time is a noise baseline signal without target characteristic peaks, proving that the detection results are negative, and indicating that the hair lysis solution of the two schemes combined with LC-MS / MS detection can accurately distinguish normal samples from drug addict samples without impurity interference.

[0048] Example 8, detection of cannabis positive samples Hair samples of suspected drug users confirmed positive for cannabis by forensic inspection were obtained, and the hair splitting solution of scheme one and scheme two was used for detection, respectively.

[0049] The detection results of scheme one are shown in Figure 8 The negative in (A) is inferred to be caused by the competition of the components with cannabis for charge in the ionization process, resulting in reduced ionization efficiency of the target and ion suppression effect. The detection results of scheme two are shown in Figure 8 (B) shows that the cannabis molecules in the hair of suspected drug users can be accurately determined to be positive, without impurity interference, which is consistent with the forensic results.

[0050] Example 9, control experiment of hair splitting solution treatment time Hair samples of suspected drug users confirmed positive for clonazepam by forensic inspection were obtained, and divided into 6 groups, each group of 5 mg, and each group of samples was labeled as A~F. Groups A~E used the hair splitting solution of scheme one for detection, and the specific operation steps were as follows: S1. Cut the hair sample to 0.5~2 mm, add 50±5 μL of the hair splitting solution to each 1 mg of hair, and vortex mix; S2. 37℃ constant temperature reaction, group A reaction time 1 minute, group B reaction time 5 minutes, group C reaction time 10 minutes, group D reaction time 15 minutes, group E reaction time 30 minutes; S3. Filter the splitting solution and perform LC-MS / MS analysis.

[0051] Group F samples were treated according to the forensic method, 50±5 μL of methanol solution was added to each 1 mg of hair, frozen and ground, ultrasonicated, filtered, and subjected to LC-MS / MS analysis.

[0052] The experimental results are shown in Table 1, samples A, B, C, D, and E all detected clonazepam, which is consistent with the detection results of sample F using physical cold grinding method, indicating that the hair splitting solution of the present application can quickly release the drug molecules in the hair. The measured clonazepam concentrations of each group were: A group 0.030 ng / mg; B group 0.045 ng / mg; C group 0.067 ng / mg; D group 0.072 ng / mg; E group 0.098 ng / mg; F group 0.087 ng / mg. It was verified by experiment that the data measured by groups D and E were closer to that of group F, i.e., the reaction time of 15~30 minutes can make the hair splitting solution release the target substance to a degree similar to the forensic results.

[0053] Example 10, control experiment of commercially available products The hair samples of Examples 1, 2 and 4 were detected by using commercially available Segue Ice Meth K three-in-one drug rapid screening plate and Segue portable drug fluorescence detector; the hair sample of Example 3 was detected by using commercially available Zhejiang Dongfang Gene Etomidate rapid screening plate and Dongfang Gene OG-G200 dry-type fluorescence immunoassay analyzer; and the results are shown in Table 1 and Table 2. Figure 9 and Table 1.

[0054] Table 1: Comparison results of commercially available products, the detection method of the present application and the forensic method

[0055] As can be seen from Table 1, the hair lysis solution combined with LC-MS / MS detection technology is good for detecting multiple drugs in the hair of suspected drug users, and the detection results are consistent with the results of the physical cold grinding method. In the hair detection experiment, when the hair lysis solution of scheme one is used for detection, the pretreatment process only takes 15 minutes. When the hair lysis solution of scheme two is used for detection, the pretreatment time is prolonged to 30 minutes (15 minutes of lysis and 15 minutes of extraction) due to the additional liquid-liquid extraction step. The traditional grinding method takes as long as 2 hours. In comparison, the two hair lysis solutions and the detection methods provided by the present application significantly improve the detection efficiency and greatly reduce the time cost required for detection, which has obvious advantages in practical application.

[0056] However, using commercially available detection tools may result in misjudgment or missed detection. In Example 1, although the Segue Ice Meth K three-in-one drug rapid screening plate and the Segue portable drug fluorescence detector detected positive for morphine, they also misjudged positive for methamphetamine. In Examples 2 and 3, the Segue Ice Meth K three-in-one drug rapid screening plate and the Segue portable drug fluorescence detector missed detection, and the detection results were negative. Only in Example 4, the Segue Ice Meth K three-in-one drug rapid screening plate and the Segue portable drug fluorescence detector correctly detected positive for methamphetamine. In comparison, the hair lysis solution and the detection method of the present application are more accurate and reliable.

[0057] It is apparent for those skilled in the art that the present application is not limited to the details of the above exemplary embodiments, but can be implemented in other concrete forms without departing from the spirit or essential characteristics of the present application. Therefore, the embodiments should be considered in all aspects as exemplary and non-limiting, and the scope of the present application is defined by the appended claims rather than the above description, and all changes falling within the meaning and range of equivalent elements of the claims are intended to be encompassed by the present application.

Claims

1. A hair lysis solution, characterized in that, The hair lysis solution contains: 0.05-1.0% by volume of ethyl phenyl polyethylene glycol, 2-18 mg / mL of dithiothreitol, and 2-20 U / mL of protease.

2. The hair lysis solution according to claim 1, characterized in that, It also includes a buffer solution selected from phosphate buffer, Tris-HCl buffer or citrate buffer.

3. The hair lysis solution according to claim 1 or 2, characterized in that, The hair lysis solution also includes 0.01 to 0.3% by volume of β-mercaptoethanol.

4. The hair lysis solution according to claim 3, characterized in that, The hair lysis solution also includes 10-40 mg / mL of sodium dodecyl sulfate.

5. The hair lysis solution according to claim 1, characterized in that, The proteases include proteinase K and / or keratinase.

6. The application of the hair lysis solution according to any one of claims 1 to 5 in the detection of drugs in human hair.

7. The application according to claim 6, characterized in that, Including the following steps: S1. Cut the hair sample into 0.5~2 mm pieces, add the hair lysis solution, and vortex mix. S2. Conduct a constant-temperature reaction; S3. Filter and perform liquid chromatography-tandem mass spectrometry analysis.

8. The application according to claim 7, characterized in that, Step S2 also includes: adding chloroform and isopropanol to the isothermal reaction product for extraction, centrifuging to remove the organic phase, drying it, and then re-dissolving it in methanol.

9. The application according to claim 7 or 8, characterized in that, The temperature of the isothermal reaction is 25~65℃.

10. The application according to claim 9, characterized in that, The chromatographic conditions for the liquid chromatography-tandem mass spectrometry analysis are as follows: Chromatographic column: Restek Allure PFP Propyl column with a length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 5 μm; Mobile phase: 5 mmol / L ammonium acetate and 0.1% formic acid buffer, eluted with acetonitrile gradient; Mass spectrometry conditions: electrospray ionization positive ion mode, multiple reaction monitoring.

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